CN104388443A - Strawberry auxin synthetic rate-limiting enzyme gene FaYUC11 and application - Google Patents

Abstract

The invention relates to an agricultural biotechnology, discloses a strawberry auxin synthetic rate-limiting enzyme gene FaYUC11 and application in adjustment of fruit sizes, and provides a nucleotide sequence and a protein sequence of the gene. A method for using the FaYUC11 to adjust the fruit sizes comprises the following steps of: constructing a virus induction silent vector of the FaYUC11 gene; and transferring the silent vector in agrobacterium, infecting young green fruits of strawberries by using a micro-injection method, and establishing a transgenosis strawberry strain silenced by the virus induction FaYUC11 gene. The content of free auxin in achenes (seeds) can be reduced through gene silencing; the growth rate of fruits in vertical and horizontal diameters is reduced; simultaneously, the hardness of fruits and the content of soluble solids are influenced; and thus, the gene disclosed by the invention has good application prospect in the aspect of adjusting and controlling the fruit sizes of strawberries, and has important significance on development of functional SNP (Single Nucleotide Polymorphism) molecular markers and breeding of high-quality large strawberries.

Description

Strawberry growth hormone synthesis rate-limiting enzyme FaYUC11 gene and application

Technical field

The present invention relates to plant biotechnology field, be specifically related to octoploid strawberry growth hormone rate-limiting enzyme FaYUC11 gene and application, and by the method for above-mentioned gene regulating strawberry fruit size.

Background technology

Strawberry (Fragaria × ananassa) belongs to Rosaceae strawberry plants, important eat one of fruit crop raw, also be the pattern species of the important functional gene research of rosaceous plant, the heredity that its fruit development is relevant and molecular biology research more and more receive publicity.Wherein, plant hormone regulation and control for fruit development and quality responses most crucial.

Strawberry fruit belongs to " pseudocarp ", and grown by holder, the fruit of phytology meaning is the seed interspersing its surface, and the latter is also called achene.As far back as eighties of last century mid-term, Nitsch just research find strawberry holder grow by the regulation and control of hormone, the growth hormone mainly synthesized in achene plays an important role (Nitsch1955).If remove the epiclinal whole achene of strawberry after pollination, holder will stop growing; And removing epiclinal Some seeds, then the part holder only containing achene is grown, the malformation fruit of last president.But, if remove seeds all on holder, then be coated on holder with artificial synthetic auxin NAA, finally can grow into normal fruit.

Growth hormone is the parahormone found the earliest in plant, so far existing nearly 80 years (Thimann andKoepfli etc. 1935).Know that growth hormone almost take part in the regulation and control of each aspect of plant life at present, comprise the growth of Various Tissues organ and morphogenesis and the response to environment.Indolylacetic acid IAA is the primary auxin in plant, and its route of synthesis may be variant because of species, organ type, etap or envrionment conditions.Recent research shows, the indolepyruvic acid approach (IPyA pathway of IAAbiosynthesis) depending on tryptophane is the primary auxin route of synthesis in various plants.Research in strawberry also shows, the synthesis of IPyA approach growth hormone is to Strawberry Growth most important (Liu etc. 2014).

The IPyA approach only comprising two-step reaction is also called TAA/YUC approach: produce indolepyruvic acid (indole-3-pyruvic acid by tryptophane under tryptophan amino transferase TAA/TAR catalysis, IPyA), IPyA is changed into IAA (Mashiguchi etc. 2011 by YUC family flavine monooxygenase afterwards; Abu-Zaitoon etc. 2012; Stepanova etc. 2011).Zhao Yunde etc. 2001 report the earliest and identify YUC from Arabidopis thalianas, for tryptophane relies on the key enzyme of growth hormone synthesis rate-limiting step.In recent years, find that the growth hormone from TAA/YUC approach is very important to growth and development of plants comprising in the various plants such as Arabidopis thaliana, paddy rice, corn, petunia, tomato and strawberry.These research supports argument, YUC extensively exists in plant, high conservative, catalyzes and synthesizes via YUC the main source that growth hormone may be growth hormone in plant.

Gene Silencing (virus-induced gene silencing, VIGS) is a kind of simple to operate, Functional identification of genes method quickly and easily.Virus vector with goal gene fragment is infected plant, vegetable cell can the threat of spontaneous identification intrusive viruses, then utilize the defense mechanism of self to resist and destroy the goal gene on virus and virus vector, thus causing goal gene at post-transcriptional level, degraded occur or even eliminate (Lange etc. 2013; Purkayastha and Dasgupta2009).There is VIGS gene silencing system in various plants for studying the reported success of corresponding gene function, particularly studying the specific gene in plant propagation organ targetedly, thus understanding growth and the quality responses mechanism of fruit.The VIGS silent carrier widely used is diplornavirus-Tobacco rattle virus (TRV) (Liu etc. 2002), successful Application (Jia etc. 2011) in strawberry.

The conventional hybridization breeding cycle of fruit tree crop is very long.Molecular Marker Assisted Selection Technology significantly can shorten breeding cycle, improves breeding efficiency.Current, molecular mark more and more highlights for the importance accelerating fruit tree conventional hybridization breeding seed selection process.Functional molecular marker (functional markers in New molecular marker, FMs), by with the functional mononucleotide polymorphism site (SNP) in the functional gene motif of phenotypic correlation based on and develop, it is advantageous that the sequence die body from controlling phenotype, with target gene close linkage, directly can apply under multiple different genetic background, can screen more effectively and accurately and follow the trail of known.Positive qualification economical character genes involved function, the exploitation and the molecular breeding work that can be functional label provide important materials and means.

The strawberry production of China deserves to be called " strawberry big country " on the total area and ultimate production, but certain gap is still there is compared with " the world strawberry is made the country prosperous " U.S., Spain etc., the ratio that self-fertile high-quality large fruited strawberry kind accounts for world's strawberry breeding is very low, and average per unit area yield is significantly lower.Disclose strawberry fruit size control molecular mechanism, the correlation function such as fruit size, weight marker development and assist-breeding are applicable to the large fruited strawberry new germ plasm of China's planting environment and consumption habit, significant.

Reference:

Nitsch JP(1955)Free auxins and free tryptophane in the strawberry.Plant Physiol 30:33-39

Thimann KV,Koepfli JB(1935)Identity of the growth-promoting and root-formingsubstances of plants.Nature 135:101–102

Liu H,Xie WF,Zhang L,Valpuesta V,Ye ZW,Gao QH,Duan K(2014)Auxin biosynthesisby the YUCCA6 flavin monooxygenase gene in woodland strawberry(Fragaria vesca).J IntegrPlant Biol 56:350-363

Stepanova AN,Yun J,Robles LM,Novak O,He W,Guo H,Ljung K,Alonso JM(2011)TheArabidopsis YUCCA1 flavin monooxygenase functions in the indole-3-pyruvic acid branch ofauxin biosynthesis.Plant Cell 23:3961-3973

Mashiguchi K,Tanaka K,Sakai T,Sugawara S,Kawaide H,Natsume M,Hanada A,YaenoT,Shirasu K,Yao H,McSteen P,Zhao Y,Hayashi K,Kamiya Y,Kasahara H(2011)The mainauxin biosynthesis pathway in Arabidopsis.Proc Natl Acad Sci U S A108:18512-18517

Abu-Zaitoon YM,Bennett K,Normanly J,Nonhebel HM(2012)A large increase in IAAduring development of rice grains correlates with the expression of tryptophan aminotransferaseOsTAR1 and a grain-specific YUCCA.Physiol Plant 146:487-499

Lange M,Yellina AL,Orashakova S,Becker A.Virus-induced gene silencing(VIGS)inplants:an overview of target species and the virus-derived vector systems.Methods Mol Biol(2013)975:1-14.

Purkayastha A,Dasgupta I.Virus-induced gene silencing:a versatile tool for discovery ofgene functions in plants.Plant Physiol Biochem.(2009)47(11-12):967-976.

Liu Y,Schiff M,Dinesh-Kumar SP.Virus-induced gene silencing in tomato.Plant J.(2002)31:777-786

Jia HF,Chai YM,Li CL,Lu D,Luo JJ,Qin L,Shen YY.Abscisic acid plays an importantrole in the regulation of strawberry fruit ripening.Plant Physiol.(2011)157:188-199

Summary of the invention:

A kind of growth hormone regulating strawberry fruit to expand is the object of the present invention is to provide to synthesize rate-limiting enzyme FaYUC11 gene and application thereof.

Technical scheme is, a kind of strawberry growth hormone synthesis rate-limiting enzyme FaYUC11 gene, containing, for example the nucleotide sequence shown in SEQ IDNo.1.Preferably, its nucleotide sequence is as shown in SEQ ID No.1.

This gene source is in octoploid strawberry, rate-limiting enzyme-YUCCA class flavine monooxygenase (FMO) in the indolepyruvic acid route of synthesis (IPyApathway of IAA biosynthesis) of encoding growth element, containing, for example the aminoacid sequence shown in SEQ ID No.2.Preferably, its aminoacid sequence is as shown in SEQ ID No.2.

Above-mentioned gene can be used for regulating and controlling indolylacetic acid (IAA) content in strawberry fruit and fruit size, and the silence of this gene not only can cause free IAA content in strawberry fruit to decline, and also makes strawberry fruit expand and is suppressed.

The present invention utilizes RT-PCR (reverse transcription PCR) and Gene Silencing VIGS (Virus-inducedgene silencing) technology to clone from octoploid strawberry and identifies a kind of new strawberry fruit size control gene FaYUC11, rate-limiting enzyme-YUCCA class flavine monooxygenase (FMO) in the indolepyruvic acid route of synthesis (IPyA pathway of IAAbiosynthesis) of this genes encoding growth hormone.The silence of this gene not only can cause free IAA content in strawberry fruit to decline, and also makes strawberry fruit expand and is suppressed.The invention provides nucleotide sequence and the protein sequence of this gene, also relate to the purposes of this gene in strawberry fruit size control simultaneously.

According to aforesaid application, the non-conservative region of strawberry growth hormone synthesis rate-limiting enzyme FaYUC11 gene is building up to the downstream of 2 × 35S promoter of pTRV2 carrier, form the two-way hairpin structure that intron (Intron) separates, transform strawberry by microinjection, and identify Transgenic Strawberry.After FaYUC11 gene silencing, strawberry fruit expand suppressed and in strawberry fruit free IAA content decline.

Make strawberry growth hormone synthesize the method for rate-limiting enzyme FaYUC11 gene silencing, be build reticent expression vector, and infect strawberry after mixing with Agrobacterium.Concrete steps are: according to the CDS sequences Design pcr amplification primer pair of strawberry growth hormone synthesis rate-limiting enzyme FaYUC11 gene, as shown in SEQ ID No.3 and No.4.

Its total length forward primer OF is: 5 '-AAAATGGAGAACAATGTGTTTGGGA-3 ';

Reverse primer OR:5 '-GGACTAGACCTCTCTTGCAGCAT-3 '.

For masterplate, utilize above-mentioned primer with octoploid " fragrant for a long time " strawberry achene cDNA, obtain FaYUC11 gene complete sequence by pcr amplification; PCR primer is cloned on pUCm-T carrier, correct through checking order.

According to strawberry YUC whole family family sequence comparison result, select FaYUC11 gene-specific region, design amplifies the gene specific primer pair of 253bp, as shown in SEQ ID No.5 and No.6.

Forward primer RiF:5 '-TTCTGCTCTCTGCCGATGAT-3 ';

Reverse primer RiR:5 '-CTCCAGTCGCAATTACCAAG-3 '.

With above the T plasmid of acquisition full-length gene for masterplate, utilize RiF and RiR primer pair, the 253bp fragment of FaYUC11 gene is obtained by pcr amplification, recombinant methods in vitro is utilized to clone Intron two ends into pBSK-in carrier (Duan etc. 2008) in two steps, define the recombinant plasmid pBSK-in-dYUC11 with 2 FaYUC11 gene fragments, wherein 2 YUC11 gene fragment directions are contrary, and centre is an intron Intron separates.To be specially: first this gene fragment is cloned on pUCm-T carrier, then will containing the pUCm-T plasmid of this FaYUC11 gene fragment through PstI and BamHI double digestion, by vitro recombination gene fragment transfer to be entered on the pUCm-T plasmid cut with like combinations enzyme; Again cut the pUCm-T plasmid containing FaYUC11 gene fragment with PstI and SalI enzyme, the gene fragment cut is entered with the linearizing pBSK-in carrier containing a FaYUC11 gene fragment of NsiI and SalI by vitro recombination transfer.

Use KpnI enzyme and SacI enzyme double digestion pBSK-in-dYUC11 recombinant plasmid further, the downstream of viral CP albumen after the two-way hairpin structure of FaYUC11 gene fragment being cloned into the 2 × 35S promoter of Tobacco rattle virus pTRV2 (Liu etc. 2002), obtains pTRV2-dYUC11 recombinant vectors and silent carrier.The Agrobacterium of RNA1 and RNA2 of mixing containing Tobacco rattle virus, adopt the little green fruit of Agrobacterium microinjection process strawberry, through observation of taking pictures continuously, RT-PCR in conjunction with TRV viral RNA 1/2 special primer and FaYUC11 detects screening, the final strawberry fruit obtaining virus induction FaYUC11 silence.

The present invention utilizes Gene Silencing method first, builds the Tobacco rattle virus silent carrier of FaYUC11, and utilizes microinjection method to be transformed into strawberry, changes fruit development, thus affects Yield of Strawberry and quality.For illustrating the hormone regulating and controlling mechanism that strawberry fruit expands in detail, there is important theory value, and can pass through based on this gene and allelic single nucleotide polymorphism research thereof, develop the functional molecular marker that fruit size is relevant, also significant in strawberry molecular breeding and the seed selection of large fruit high-quality strawberry.

Accompanying drawing explanation

Fig. 1 is virus induction silent carrier (pTRV2-dYUC11 carrier) collection of illustrative plates of constructed target gene in the embodiment of the present invention 1.

Fig. 2 is the RT-PCR qualification result of virus-positive strain in the embodiment of the present invention 3.Adopt TRV2 special primer to carrying out the detection of pcr amplification rear electrophoresis, wherein swimming lane 1 is the fruit of octoploid strawberry " fragrant for a long time ", swimming lane 2 be CK1 not containing the of a specified duration fragrant fruit that the TRV Agrobacterium of target gene fragment is injected, swimming lane 3 be RiYUC11 target gene silent carrier Agrobacterium injection of a specified duration fragrant fruit.

Fig. 3 is the qRT-PCR qualification result of FaYUC11 gene silencing in the embodiment of the present invention 3, is the expression of strawberry growth hormone synthetic gene FaYUC11 in contrast (CK1) the vanilla certain kind of berries of a specified duration and 2 Gene Silencing strains.Ordinate zou is that real-time PCR (real-time fluorescence quantitative PCR) detects the relative expression quantity (wild-type relative to non-injecting virus silent carrier) of this gene in contrast CK1 or Gene Silencing strain.

Fig. 4 is fruit phenotype analytical after Gene Silencing in the embodiment of the present invention 4, for contrast (CK1 does not transform containing the viral empty carrier that target gene inserts) and the form of 2 Gene Silencing strain (RiYUC11-2 and RiYUC11-3) fruit process after 34 days.Compared with the control, Gene Silencing Fruit is obviously suppressed, and it is normal to contrast Fruit.

Fig. 5 is Gene Silencing strain and the content difference contrasting free I AA in strawberry achene in the embodiment of the present invention 4.Ordinate zou shows the IAA content in the strawberry achene measured by GC-MS method.

Fig. 6 is the difference of the fruit longitudinal and transverse footpath growth ratio of the reticent strain of virus induction and contrast in the embodiment of the present invention 4.Before and after ordinate zou display microinjection, the change of longitudinal and transverse footpath is compared to the ratio before injection.

Embodiment

Following examples are only not used in for illustration of the present invention and limit range of application of the present invention.

The separation of embodiment 1FaYUC11 gene and Gene Silencing vector construction

According to diploid forest strawberry YUC whole family race's gene sequence information (Liu etc. 2014), in octoploid planting strawberry fruit (point holder and achene), resolve the Expression pattern of whole family race gene.Found that FaYUC11 is this family uniquely high expression level member in achene, and its Expression pattern is consistent with growth hormone accumulation dynamic pattern in achene.

According to forest strawberry homogenic sequences Design pcr amplification primer, forward primer OF:

5 '-AAAATGGAGAACAATGTGTTTGGGA-3 '; Reverse primer OR:

5 '-GGACTAGACCTCTCTTGCAGCAT-3 ', respectively as shown in SEQ ID No.3 and 4.

With wild-type fragrant strawberry fruit cDNA of a specified duration for masterplate, full formula gold high-fidelity enzyme (from Beijing TransGene company) is adopted to carry out pcr amplification with above-mentioned amplimer, obtain FaYUC11 complete sequence, its nucleotide sequence is as shown in SEQ ID No.1, and the aminoacid sequence of albumen coded by it is as shown in SEQ ID No.2.

PCR reaction is carried out according to TranStart polymeric enzymatic amplification system and response procedures.PCR primer being connected is cloned on pUCm-T carrier, obtains the identical sequence with goal gene, as SEQ IDNo.1 through order-checking qualification.

Afterwards, again according to the sequence alignment result of the full family member of vanilla certain kind of berries YUC of a specified duration, choose the gene-specific region of holding by 5 ', devise interference primer pair, forward primer: 5 '-TTCTGCTCTCTGCCGATGAT-3 ' and reverse primer: 5 '-CTCCAGTCGCAATTACCAAG-3 ', respectively as shown in SEQ ID No.5 and 6.

With acquisition full-length gene plasmid for masterplate, utilize and above-mentionedly designed interfere primer pair, pcr amplification obtains the gene fragment of 253bp.First this gene fragment is cloned on pUCm-T carrier, and then the intron sequence two ends of pBSK-in carrier are transferred to through twice vitro recombination, be integrated in viral RNA 2 plasmid pTRV2 finally by KpnI and SacI double digestion point, obtain pTRV2-dYUC11 Gene Silencing carrier.Vector map is shown in Fig. 1.

Wherein, the building process of intermediate carrier pBSK-in-dYUC11 is: first by the pUCm-T plasmid containing FaYUC11 gene fragment (253bp) through PstI and BamHI double digestion, by vitro recombination, gene fragment transfer is entered on the pUCm-T plasmid (Shanghai Sheng Gong biotechnology company limited) cut with like combinations enzyme.Afterwards, again cut the pUCm-T plasmid containing FaYUC11 gene fragment with PstI and SalI enzyme, the gene fragment cut is entered with the linearizing pBSK-in carrier containing a target gene fragment of NsiI and SalI by vitro recombination transfer, just obtain the recombinant plasmid pBSK-in-dYUC11 containing 2 FaYUC11 gene fragments, wherein 2 YUC11 gene fragment directions are contrary, and centre is separated by an intron (Intron).

The acquisition of embodiment 2 Gene Silencing strawberry

Use KpnI enzyme and SacI enzyme double digestion pBSK-in-dYUC11 recombinant plasmid further, the downstream of viral CP albumen after the two-way hairpin structure of FaYUC11 gene fragment being cloned into the 2 × 35S promoter of Tobacco rattle virus pTRV2 (Liu etc. 2002), obtains pTRV2-dYUC11 recombinant vectors.The Agrobacterium of RNA1 and RNA2 of mixing containing Tobacco rattle virus.

By the GV3101 Agrobacterium inoculation containing the reticent expression vector of target gene virus induction and empty pTRV1 and pTRV2 carrier in containing 50 μ g/mL Kan, 28 DEG C of overnight incubation in the 5mL YEP substratum of 10 μ g/mL Rif and 50 μ g/mL Gen, next day is inoculated in inducing culture (YEP according to the ratio of 1:50, containing 10mM/L MES, 20 μMs/L AS, 50 μ g/mL Kan, 10 μ g/mLRif and 50 μ g/mL Gen) in, 28 DEG C are shaken bacterium to logarithmic phase (OD value 0.6-0.8), the centrifugal 10min of 5000r/min collects thalline, again with infecting damping fluid (containing 10mM/L MES, 100 μMs/L AS, 10mM/L MgCl ₂) resuspended thalline, and regulate bacterial concentration to OD value between 1.2-1.5, for inoculating strawberry fruit (with " fragrant for a long time " the little green fruit of strawberry in intelligent artificial greenhouse for material) after 28 DEG C of placement 3h.Then the GV3101 bacterium liquid containing pTRV1 is pressed 1:1 volume with the GV3101 bacterium liquid containing pTRV2-dYUC11 to mix, use microinjection method by the bacterium liquid of mixing, inject from fruit top, the temperature that controls environment, at 25-28 DEG C, ensures that virus effectively infects.

The qualification of embodiment 3 Gene Silencing strawberry

Whether successfully infecting strawberry to detect Tobacco rattle virus, extracting the strawberry fruit total serum IgE of the virus induction silent carrier of the FaYUC11 of wild-type and injecting virus initial carrier and restructuring respectively, utilize random primer to carry out reverse transcription synthesis cDNA.

There is provided the special primer of Tobacco rattle virus RNA2 to carry out RT-PCR reaction according to 2011 documents such as Jia, carry out the strawberry fruit that identifying virus infects.Carry out electrophoresis detection to PCR primer, found that, the strawberry fruit that the recombinant viral vector of viral initial carrier and FaYUC11 infects can amplify viral specific band (as Fig. 2), and without this object band in wild-type strawberry.Tentatively determine, Tobacco rattle virus has infected strawberry fruit by microinjection method.

Further, in the fragment downstream of the viral silent carrier of selected structure, the special primer pair that design FaYUC11 is new.Forward primer: 5 '-GGAAAGGTGAAAAGGGCGT-3 '; Reverse primer:

5’-GACCTCTCTTGCAGCATTAC-3’。

By the effect of RT-PCR in transcriptional level identifying virus induced gene silence.Get two to verify by the FaYUC11 gene silencing strain of recombinant virus induction, represent with RiYUC11-2 and RiYUC11-3 respectively.

Result shows, compared to the fruit of injection containing protovirus plasmid Agro-Bacterium, the accumulating level of transcription product in various degree that there occurs FaYUC11 in the fruit of part microinjection containing the reticent plasmid Agro-Bacterium of virus induction of FaYUC11 restructuring declines, as Fig. 3.

Embodiment 4 Gene Silencing strawberry phenotype analytical

Protovirus is infected strawberry fruit to compare with the strawberry fruit of wild-type with developmental stage, after finding the Agrobacterium injection containing original TRV virus, slight depression is arranged at fruit top, and Fruit and grown form are not subject to obvious impact in addition.Further phenotype analytical finds, the strawberry fruit of injection recombinant viral vector Agrobacterium expands and is suppressed, and part fruit does not almost become large through the growth of month, is still in the little green fruit stage, the suppression suffered by part Fruit is had to want slight, as Fig. 4 in addition.

Measure the IAA in strawberry fruit seed (achene) by GC-MS method, obviously can find out that the strawberry fruit of virus induction FaYUC11 silence not only expands and be suppressed, growth hormone content also obviously declines, as Fig. 5.Measure further by fruit longitudinal and transverse demeter, compared with fruit size data before containing viral Agrobacterium with microinjection, obviously can find out that the strawberry fruit of the virus induction silent carrier injection of restructuring FaYUC11 is less than the longitudinal and transverse demeter growth ratio of protovirus vector injection fruit, as Fig. 6.

Result proves, this gene silencing can cause free growth hormone content in fruit achene (seed) to decline, and fruit longitudinal and transverse demeter rate of increase reduces, and meanwhile, have impact on the hardness of fruit and soluble solid content.This gene has good application prospect in strawberry fruit size control, for the exploitation of functional SNP marker and the seed selection of high-quality large fruited strawberry significant.

Although above the present invention is described in detail with a general description of the specific embodiments, on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements done without departing from theon the basis of the spirit of the present invention, all belong to the scope of protection of present invention.

Claims (9)

1. strawberry growth hormone synthesis rate-limiting enzyme FaYUC11 gene, is characterized in that, containing the nucleotide sequence shown in SEQ ID No.1.

2. strawberry growth hormone synthesis rate-limiting enzyme FaYUC11 gene, it is characterized in that, its nucleotide sequence is as shown in SEQID No.1.

3. the application of strawberry growth hormone synthesis rate-limiting enzyme FaYUC11 gene in regulating fruit size described in claim 1 or 2.

4. described in claim 1 or 2, strawberry growth hormone synthesis rate-limiting enzyme FaYUC11 gene is used for the content of indolylacetic acid in regulating fruit.

5. the raw phytosynthesis rate-limiting enzyme of strawberry, is characterized in that, containing the aminoacid sequence shown in SEQ ID No.2.

6. the raw phytosynthesis rate-limiting enzyme of strawberry described in claim 5, it is characterized in that, aminoacid sequence is as shown in SEQ IDNo.2.

7. the raw phytosynthesis rate-limiting enzyme of strawberry described in claim 5, is characterized in that, by genes encoding described in claim 1 or 2.

8. the method for claim 1 or described strawberry growth hormone synthesis rate-limiting enzyme FaYUC11 gene silencing, is characterized in that, build reticent expression vector, and infect strawberry after mixing with Agrobacterium.

9. the method for strawberry growth hormone synthesis rate-limiting enzyme FaYUC11 gene silencing described in claim 8, it is characterized in that, the construction process of described reticent expression vector is:

With the sequence of SEQ ID No.5 and No.6 for primer, FaYUC11 full length gene sequence is template, obtains the FaYUC11 gene fragment of 253bp;

First this gene fragment is cloned on pUCm-T carrier, then will containing the pUCm-T plasmid of this FaYUC11 gene fragment through PstI and BamHI double digestion, gene fragment transfer to be entered on the pUCm-T plasmid cut with like combinations enzyme by vitro recombination;

Again cut the pUCm-T plasmid containing FaYUC11 gene fragment with PstI and SalI enzyme, the gene fragment cut is entered with the linearizing pBSK-in carrier containing a FaYUC11 gene fragment of NsiI and SalI by vitro recombination transfer, obtain the recombinant plasmid pBSK-in-dYUC11 containing 2 FaYUC11 gene fragments, wherein 2 YUC11 gene fragment directions are contrary, and centre is an intron separates;

Use KpnI enzyme and SacI enzyme double digestion pBSK-in-dYUC11 recombinant plasmid further, the downstream of viral CP albumen after the two-way hairpin structure of FaYUC11 gene fragment is cloned into the 2 × 35S promoter of Tobacco rattle virus pTRV2, obtain pTRV2-dYUC11 recombinant vectors, be reticent expression vector.